Mempro™ Detergent-Free Pentameric Ligand-Gated Ion Channel Production

The pLGIC, a single neurotransmitter receptor superfamily, mediates fast chemical transmission by converting nerve signals into an electrical response. It has provent that pLGICs are a group of transmembrane ion channels that open to allow ions such as Na+, K+, Ca2+, or Cl− to pass through the cell membrane, which are composed of five homologous subunits. And theses subunits consist of at least two different domains, including a transmembrane domain which forms the channel gate and the ion selective filter, and an extracellular domain which includes the ligand binding location (an allosteric binding site). In addition, pLGICs are divided into three classes: Cys-loop receptors, Ionotropic glutamate receptors and ATP-gated channels. Traditional approaches for pLGIC purification need extraction from the membrane into detergent micelles. However, detergent can induce conformational change on membrane proteins. Our alternative detergent-free expression system can best reserve the native conformation of pLGICs for further structural studies.

Nanodiscs are the self-assembly system to stabilize membrane proteins removed from the membrane by membrane scaffold proteins (MSPs) in no need of detergent. Creative Biostructure can reconstitute pLGICs into nanodiscs in detergent-free process: Expressing the pLGICs in a cell-free system with the addition of pre-assembled nanodiscs.

Mempro™ pLGIC production using amphipols

The amphipathic amphipols have the ability to “trap” around transmembrane regions of proteins, allowing them to stay folded. Creative Biostructure can utilize A8-35 to solubilize pLGICs during purification step.